Summary SHMT2 regulates one-carbon transfer reactions essential for amino acid and nucleotide metabolism, using PLP as a cofactor. Apo SHMT2 exists as a dimer with unknown functions, whereas PLP binding stabilizes the active, tetrameric state. SHMT2 also promotes inflammatory cytokine signaling by interacting with the BRISC deubiquitylase (DUB) complex, although it is unclear if this function relates to metabolism. We reveal the cryo-EM structure of human BRISC-SHMT2 complex at 3.8 Å resolution. The BRISC complex is a U-shaped dimer of four subunits and SHMT2 sterically blocks the BRCC36 active site and inhibits DUB activity. Only the inactive SHMT2 dimer, but not the active, PLP-bound tetramer binds and inhibits BRISC. BRISC mutations that disrupt SHMT2 binding impaired type I interferon signaling in response to inflammatory stimuli. Intracellular PLP levels regulated BRISC-SHMT2 interaction and inflammatory cytokine responses. These data reveal a new mechanism of metabolite regulation of DUB activity and inflammatory signaling.

The ubiquitin (Ub) proteasome system and Ub signalling networks are crucial to cell biology and disease development. Deubiquitylases (DUBs) control cell signalling by removing mono-Ub and polyubiquitin chains from substrates. DUBs take part in almost all processes that regulate cellular life and are frequently dysregulated in disease. We have catalogued 99 currently known DUBs in the human genome and sequence conservation analyses of catalytic residues suggest that 11 lack enzyme activity and are classed as pseudo-DUBs. These pseudoenzymes play important biological roles by allosterically activating catalytically competent DUBs as well as other active enzymes. Additionally, pseudoenzymes act as assembly scaffolds of macromolecular complexes. We discuss how pseudo-DUBs have lost their catalytic activity, their diverse mechanisms of action and their potential as therapeutic targets. Many known pseudo-DUBs play crucial roles in cell biology and it is likely that unstudied and overlooked pseudo-DUB genes will have equally important functions.